Diclofenac sodium 75 mg/Misoprostol 200 mcg is a combination medication commonly used for the treatment of arthritis and to prevent ulcers in people who are at risk while taking nonsteroidal anti-inflammatory drugs (NSAIDs). Assay testing for this combination product involves analyzing both diclofenac sodium and misoprostol to ensure their proper quantities and quality in the finished product.
SOP for Finished Product Analysis of Diclofenac sodium 75 mg/Misoprostol 200 mcg By UV and HPLC Method ensures that the diclofenac sodium 75 mg/Misoprostol 200 mcg tablets contain the correct amounts of both active ingredients, which is important for their effectiveness and safety. It helps to maintain the quality and efficacy of the medication and ensures compliance with regulatory requirements and standards.
PURPOSE:
To describes the procedure for analysis at in-process and finished stage of Diclofenac sodium 75 mg/Misoprostol 200 mcg Tablet.
SCOPE:
This SAP gives detailed outline for the finished product analysis of Diclofenac sodium 75 mg/Misoprostol 200 mcg Tablet and will cover In process testing activities on physical, chemical & instrumental basis.
RESPONSIBILITY:
QC Analyst is responsible for physical / chemical testing and preparing standard analytical procedure.
It is the responsibility of QC Manager to assist and ensure Testing Procedure as per SAP and to make certain that this SAP is followed in its entirety, reviewed regularly and revised as necessary.
REFERENCE:
USP 44, NF 39
MATERIAL AND EQIUPMENT:
- HPLC
- Dissolution apparatus
- Friability apparatus
- Hardness apparatus
- Disintegration Tester
- Vernier Caliper
- Analytical Balance
- pH meter
- Moisture analyzer
- Mortar and pestle
- Spatula
- Filter Paper
- Magnetic Stirrer & Hot Plate
- Sonicator
- Vacuum Pump
- Glassware
- Ammonium Acetate
- Acetonitrile HPLC Grade
- Methanol HPLC Grade
- Distilled water
- Phosphoric acid
- Monobasic Potassium phosphate
PROCEDURE:
FINAL MIX
Description:
White powder.
Procedure: Take 2.0 g of the test sample in a watch glass and observe visually with black background. Check the appearance of color, nature and any visible foreign particles.
Identification: (By HPLC)
SOP for Finished Product Analysis of Diclofenac sodium 75 mg/Misoprostol 200 mcg By UV and HPLC Method
Misoprostol
Diluent: Methanol and water (4:1)
Standard solution: 16 ug/mL of USP Misoprostol RS in Diluent. [NOTE-If outer misoprostol layers of the Tablets contain hypromellose, the Standard solution should also contain hypromellose at the same concentration as in the Sample solution.]
Sample solution: Gently break up one by one a quantity of Tablets equivalent to 0.4 mg of misoprostol, and remove the inner diclofenac layers. [Note-Keep the diclofenac layers for Identification A, Diclofenac sodium.]Transfer the outer misoprostol layers to a 25-mLvolumetric flask. Add about 15 mL of Diluent, shake for 30min, dilute with Diluent to volume, and mix well. Transfer a portion of the solution into a glass centrifuge tube, and centrifuge for 10 min under refrigerated conditions (10°).Use the supernatant.
Blank: Diluent
Cell: 1 cm
Acceptance criteria: Meet the requirements
Diclofenac sodium
Standard solution: 0.1 mg/mL of USP Diclofenac Sodium RS in methanol
Sample solution: Transfer the diclofenac inner layers reserved from Identification A, Misoprostol, to a 100-mLvolumetric flask. Add about 60 mL of methanol, shakefor10 min, dilute with methanol to volume, and mix well. Further dilute a suitable volume of the solution to obtain a solution containing about 0.1 mg/mL of diclofenac sodium, based on the label claim. Pass a portion of the solution through a polytetrafluoroethylene (PTFE) with glass microfiber (GMF) filter of 0.45-~m pore size.’ Discard the first few milliliters of the filtrate, and use the filtrate.
Blank: Methanol
Cell: 0.05 cm
Acceptance criteria: Meet the requirements
Misoprostol: The retention time of the misoprostol peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Misoprostol.
Diclofenac sodium: The retention time of the diclofenac peak of the Sample solution corresponds to that of the Standard solution, as obtained in the Assay for Diclofenac Sodium.
Loss on Drying: (By Moisture Analyzer)
Procedure:
Turn on instrument 20 minute before the test. Take 2 g powder spread uniformly on Moisture analyzerplate. Close the lid and press start button.
NMT 5.0% when determined by LOD Apparatus, Use 2.0 g of powder.
Assay By HPLC
MISOPROSTOL
Buffer: Prepare 0.025 M monobasic potassium phosphate, pH 6.5, as follows. Adjust a solution containing 3,4 g/L of monobasic potassium phosphate in water with 1 N sodium hydroxide to a pH of 6.5.
Mobile phase: Acetonitrile and Buffer (45:55)
Standard solution: 0.01 mg/mL of USP Misoprostol RS in Mobile phase, using sonication as needed
Sample solution: Nominally 0.01 mg/mL of misoprostol prepared as follows. Using a quantity of Tablets equivalent to 5 mg of misoprostol, place 1 Tablet at a time on its edge inside a well-folded piece of weighing paper. Tap very carefully the edge of the Tablet with a pestle to separate the Tablet into the outer and inner layers. Remove the inner core containing diclofenac sodium. Transfer the outer portions of the Tablets, containing misoprostol, into a500-mL volumetric flask containing a magnetic stir bar, and add 250 mL of acetonitrile. Stir the flask for 1 h. Add 150mL of water, and stir for an additional 30 min or until the Tablets are completely disintegrated. Remove the stir bar,
rinse it inside the flask with water, dilute with water to volume, and mix well. Transfer a portion of the solution in to a glass centrifuge tube, and centrifuge for 10 min under refrigerated conditions (10°). Use the supernatant.
Chromatographic system
(See Chromatography (621), System Suitability.)
Mode: LC
Detector: UV 200 nm
Column: 4.6-mm x 25-cm; 5um packing L10
Temperatures: Autosampler: 10° and Column: 35°
Flow rate: 1.0 mL/min
Injection volume: 80 uL
System suitability
Sample: Standard solution
Suitability requirements
Tailing factor: NMT 2
Relative standard deviation: NMT 2.0% Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of the labeled amount of misoprostol (C22H38O5) in the portion of Tablets taken:
Result= (Ru/Rs) x (Cs/Cu) x 100
Ru=peak response from the Sample solution
Rs= peak response from the Standard solution
Cs= concentration of USP Misoprostol RS in the Standard solution (mg/mL)
Cu=nominal concentration of misoprostol in the Sample solution(mg/mL)
Limit: 90%-110%of the labeled amount
DICLOFENAC SODIUM
Buffer: Mix equal volumes of 0.01 M phosphoric acid and0.01 M monobasic sodium phosphate. If necessary, adjust with additional portions of the appropriate component to a pH of 2.5.
Mobile phase: Methanol and Buffer (70:30) Diluent: Methanol and water (70:30)
System suitability solution: 20 ug/mL of diethyl phthalate,8 ug/mL of USP Diclofenac Related Compound A RS, and0.75 mg/mL of USP Diclofenac Sodium R in Diluent.
Standard solution: 0.75 mg/mL of USP Diclofenac Sodium RS in Diluent, using sonication as needed
Sample stock solution: Transfer a quantity of Tablets, equivalent to 1500 mg of diclofenac sodium, into a1000 mL volumetric flask containing a magnetic stir bar. Add 700 mL of Diluent, and stir for 60 min or until the tablets are completely disintegrated. Remove the stir bar, rinse it with Diluent, and sonicate the sample for 15 min. Allow the sample to cool to room temperature, dilute with Diluent to volume, and mix well.
Sample solution: Nominally 0.75 mg/mL of diclofenac sodium prepared as follows. Transfer10.0 mL of the Sample stock solution into a 20-mL volumetric flask, and dilute with Diluent to volume. Pass a portion of the solution through a PTFE with GMF filter of 0,45um pore size, discarding the first few milliliters of the filtrate.
Chromatographic system
(See Chromatography (621), System Suitability.)
Mode: LC
Detector: UV 254 nm
Column: 4.6-mm x 25-cm; 5um packing L7
Flow rate: 1.0 mL/min
Injection volume: 10 uL
System suitability
Samples: System suitability solution and Standard solution
[NOTE-The relative retention times for diethylphthalate, diclofenac related compound A, and diclofenac are about 0.6, 0.7, and 1.0, respectively.]
Suitability requirements
Resolution: NLT 2.2 between the diethyl phthalate and diclofenac related compound A peaks; NLT 6.5 between the diclofenac related compound A and diclofenac peaks, System suitability solution
Tailing factor: NMT 2, Standard solution
Relative standard deviation: NMT 2.0%, Standard Solution
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of the labeled amount of diclofenac sodium (C14H10CI2NNaO2) in the portion of tablets taken:
Result= (Ru/Rs) x (Cs/Cu) x 100
Ru=peak response from the Sample solution
Rs= peak response from the Standard solution
Cs=concentration of Diclofenac Sodium RS in the Standard solution (mg/mL)
Cu=nominal concentration of Diclofenac Sodium in the Sample solution(mg/mL)
Calculations:
Average Sample AUC x Standard Concentration x Ave. Weight of Tablet x Potency
Average Standard AUC x Sample Concentration x Label Claim
(Limit for Diclofenac Sodium: 90%-110%of the labeled amount)
ALTERNATE METHOD BY UV
Standard Solution:
Take 75 mg of Diclofenac Sodium WS dissolved in 0.1N NaOH and make up the volume to 100mL with 0.1N NaOH. Shake, and sonicate for 5 minutes to facilitate dissolution
Take 2 mL from standard stock solution and make up the volume to 100 mL with 0.1N NaOH.
Sample solution:
Take weight of finally grinded granules equivalent to 75mg of Diclofenac Sodium and dissolved in 0.1N NaOH and make up the volume to 100mL with 0.1N NaOH. Shake by mechanical mean for 10 minutes to disperse and allow any insoluble matter to settle.
Pass a portion of the sample stock solution through a suitable filter of 0.45um pore size.
Transfer the 2 mL of filtrate to a 100 mL volumetric flask make up volume with 0.1N NaOH. Take reading at UV spectrophotometer at 276 nm using 0.1N NaOH as blank.
Calculations:
Sample Absorbance x Standard Concentration x Ave. Weight of Tablet x Potency
Standard Absorbance x Sample Concentration x Label Claim
(Limit for Diclofenac Sodium: 90%-110%of the labeled amount)
Hardness Test & Dimensions:
Perform the hardness test on 10 tablets and take the average. Power on the instrument and regulate zero adjustment and preset the ‘’HARDNESS’’ and “THICKNESS” mode. Place the tablet on the test plate, begin testing and read the hardness thickness and diameter. Clean the shattered tablet and print out test results, perform the test on 10 tablets and take the average.
Hardness Standard Limit: NMT 10 Kg
Friability Test:
For tablets with a unit mass equal to or less than 650 mg, take a sample of whole tablets corresponding to 6.5 g. For tablets with a unit mass of more than 650 mg, take a sample of 10 whole tablets. The tablets should be carefully dedusted prior to testing. Accurately weigh the tablet sample, and place the tablets in the drum. Rotate the drum 100 times, and remove the tablets. Remove any loose dust from the tablets as before, and accurately weigh. Generally, the test is run once. If obviously cracked, cleaved, or broken tablets are present in the tablet sample after tumbling, the sample fails the test. If the results are doubtful or if the weight loss is greater than the targeted value, the test should be repeated twice and the mean of the three tests determined. A maximum mean weight loss from the three samples of not more than 1.0% is considered acceptable for most products.
% FRIABILITY= W1-W2 x 100/W1
Dissolution Test:
Misoprostol:
USP Apparatus: USP Apparatus II Paddle
Medium: Water; 500 mL, deaerated
Speed: 50rpm
Volume: 500mL
Recommended Sampling Time: 20 minute
Buffer: Prepare as directed in the Assay for Misoprostol.
Mobile phase: Acetonitrile and Buffer (42:58).
Standard stock solution: Transfer 4 mg of USP Misoprostol RS in to a 100 mL volumetric flask, add 20 mL of acetonitrile, arid shake for about 15 min. If the outer misoprostol layers of the Tablets contain hypromellose, add a suitable amount of hypromellose to the flask to achieve the same final concentration of hypromellose in the Standard solution as expected in the Sample solution.
Add 20 mL of water, and sonicate for about 2 min. Add water up to the neck of the flask, and allow the solution to cool to room temperature before the final dilution to volume.
Standard solution: About 0.0004 mg/mL of USP Misoprostol RS prepared as follows. Dilute 2.0 ml of the Standard stock solution with Medium to 200 ml.
Sample solution: Pass a portion of the solution under test through a suitable filter of 10um pore size.
Chromatographic system:
Proceed as directed in the Assay for Misoprostol, except for Injection volume.
Injection volume: 200 ul
System suitability
Sample: Standard solution
Suitability requirements
Tailing factor: NMT 2
Relative standard deviation: NMT 5.0%
Analysis
Samples: Standards solution and Sample solution
Calculate the percentage of the labeled amount of misoprostol (C22H38O5) dissolved:
Result=(Ru/Rs) x (Cs/L) x V x 100
R u = peak response from the Sample solution
R s =peak response from the Standard solution
Cs =concentration of USP Misoprostol RS in the Standard solution (mg/mL)
L = label claim for misoprostol (mg/Tablet)
V =volume of Medium, 500 ml
Tolerances: NLT 75% (Q) of the labeled amount of misoprostol (C22H38O5) is dissolved.
Diclofenac sodium
Proceed as directed in Dissolution (711),
Apparatus 1 and Apparatus 2, Delayed-Release Dosage Forms, Method A Procedure.
Acid stage medium: 0.1 N hydrochloric acid; 750 mL, deaerated
Buffer stage medium: After 2 h, add 250 rnl, of 0.2 M tribasic sodium phosphate to the Acid stage medium and, if needed, adjust with either 2 N hydrochloric acid or 2 N sodium hydroxide to a pH of 6.8.
Apparatus 2: 100 rpm
Times: 2 h for Acidstage;45 min for Buffer stage
Buffer: 0.025 M monobasic potassium phosphate buffer with a pH of 3.0 prepared as follows; Adjust a solution containing 3.4 g/L of monobasic potassium phosphate in water with phosphoric acid to a pH of 3.0.
Mobile phase: Acetonitrile and Buffer (60:40)
Standard stock solution: 0.68 mg/mL of USP Diclofenac Sodium RS, first dissolved in 0.1 N sodium hydroxide using about 10% of the final volume, and then diluted with water to volume
Chromatographic system
(See Chromatography (621), System Suitability.)
Mode: LC
Detector: UV 276 nm
Column: 4.6-mm x 15-cm; 5 um packing L7
Flow rate: 1.0 mL/min
Injection volume: 10 uL
Acid stage
Acid stage standard solution: 13.6 ug/mL of USP Diclofenac Sodium RS prepared as follows. Transfer 2.0mL of the Standard stock solution to a 100 mL volumetric flask, and dilute with a mixture of 0.1 N hydrochloric acid and 5 N sodium hydroxide (900:20) to volume.
Acid stage sample solution: Run the test in Acid stage medium for 2 h. Withdrawal 10-mLaliquot, transfer it to a flask containing 1.0 mL of 1 N sodium hydroxide, and mix well. Pass a portion of this solution through a suitable filter of 10um pore size.
System suitability
Sample: Acid stage standard solution
Suitability requirements
Tailing factor: NMT 2
Relative standard deviation: NMT 2.0%
Analysis
Samples: Acid stage standard solution and Acid stage sample solution
Calculate the percentage of the labeled amount of diclofenac sodium (C14H10Cl2NNaO2) dissolved during the Acid stage:
Result = (Au/As) x Cs x V x D x (1/L) X 100.
Au =peak response from the Acid stage sample solution
As = peak response from the Acid stage standard solution
Cs= concentration of USP Diclofenac Sodium RS in the Acid stage standard solution (mg/mL)
L = label claim for diclofenac sodium (mg/Tablet)
V = volume of Acid stage medium, 750 mL
D = dilution factor for the Acid stage sample solution,1.1
Tolerances: NMT 10% of the labeled amount of diclofenac sodium (C14H10Cl2NNaO2) is dissolved. The percentage of the labeled amount of diclofenac sodium dissolved at the time specified conforms to Dissolution
Buffer stage
Buffer stage standard solution: 13.6 ug/mL of USP Diclofenac Sodium RS prepared as follows. Transfer 2.0mL of the Standard stock solution to a 100 mL volumetric flask, and dilute with Buffer stage medium to volume.
Buffer stage sample solution: Pass a portion of the solution under test through a suitable filter of 10umpore size.
System suitability
Sample: Buffer stage standard solution
Suitability requirements
Tailing factor: NMT 2
Relative standard deviation: NMT 2.0%
Analysis
Samples: Acid stage sample solution, Buffer stage standard solution, and Buffer stage sample solution
Calculate the percentage of the labeled amount of diclofenac sodium (C14H10Cl2NNaO2) dissolved during the Buffer stage:
Result=(Au/As) x (Cs /L)x (V- Vs) x 100
A u = peak response from the Buffer stage sample solution
As =peak response from the Buffer stage standard solution
Cs = concentration of USP Diclofenac Sodium RS in the Buffer stage standard solution (mg/mL)
L = label claim for diclofenac sodium (mg/Tablet)
V = volume of Buffer stage medium, 1000 mL
Vs = volume of the Acid stages ample solution, 10 mL
Tolerances: NLT 75% (Q) of the labeled amount of diclofenac sodium (C14H10Cl2NNaO2) is dissolved. The percentage of the labeled amount of diclofenac sodium dissolved at the time specified conforms to Dissolution